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This study focuses on 1 H NMR spectroscopic analyses of urine, which in comparison to blood is a non-invasive biofluid to access, making it a more attractive choice for large-scale biological sampling in children. The influence of the sample collection time-point of the day on the metabolic phenotype was examined, followed by assessment of the analyte detectability and quantification, as well as the likely sources of short term variability within and between children. We confirm the high analytical reproducibility and robustness of NMR-based urinary metabolic phenotyping and illustrate the benefits of pooling spot urines when seeking the stable component of the metabolome.

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